Automated otolith image classification with multiple views: an evaluation on Sciaenidae.

Combined multiple 2D views (proximal, anterior and ventral aspects) of the sagittal otolith are proposed here as a method to capture shape information for fish classification. Classification performance of single view compared with combined 2D views show improved classification accuracy of the latter, for nine species of Sciaenidae. The effects of shape description methods (shape indices, Procrustes analysis and elliptical Fourier analysis) on classification performance were evaluated. Procrustes analysis and elliptical Fourier analysis perform better than shape indices when single view is considered, but all perform equally well with combined views. A generic content-based image retrieval (CBIR) system that ranks dissimilarity (Procrustes distance) of otolith images was built to search query images without the need for detailed information of side (left or right), aspect (proximal or distal) and direction (positive or negative) of the otolith. Methods for the development of this automated classification system are discussed.

Tetracycline marks visible in Baltic cod Gadus morhua otoliths stored for 40 years.

Baltic cod Gadus morhua otoliths marked with oxytetracycline (OTC) in 1974-1975 had been sectioned and the sections stored in paper envelopes at room temperature in dry, dark conditions. After 40 years of storage, the historic OTC marks were still clearly visible showing that OTC is suitable for long-term chemical marking of otoliths. They were, however, noticeably paler than marks in recently recaptured and archived Baltic G. morhua otoliths, chemically tagged with tetracycline at similar dosage to the historic otoliths.

Three-dimensional rendering of otolith growth using phase contrast synchrotron tomography.

A three-dimensional computer reconstruction of a plaice Pleuronectes platessa otolith is presented from data acquired by the Diamond Light synchrotron, beamline I12, X-ray source, a high energy (53-150 keV) source particularly well suited to the study of dense objects. The data allowed non-destructive rendering of otolith structure, and for the first time allows otolith annuli (internal ring structures) to be analysed in X-ray tomographic images.

Detection of human utricular otoconia degeneration in vital specimen and implications for benign paroxysmal positional vertigo.

Otoconia are assumed to be involved in inner ear disorders such as benign paroxysmal positional vertigo (BPPV). Up to now, the distinct structure and morphology of intact and degenerate human utricular otoconia has been only poorly investigated on vital specimen. In this study, human otoconia were obtained from the utricle in five patients undergoing translabyrinthine vestibular schwannoma surgery. Specimens were examined by environmental scanning electron microscopy. Intact and degenerate otoconia as well as fracture particles of otoconia and bone were analyzed by energy dispersive X-ray microanalysis (EDX) and powder X-ray diffraction (XRD). Intact otoconia reveal a uniform size showing characteristic symmetry properties. Degenerative changes can be observed at several stages with gradual minor and major changes in their morphology including fragment formation. EDX analyses reveal the characteristic chemical composition also for otoconia remnants. XRD shows that intact and degenerate otoconia as well as remnants consist of the calcite modification. In conclusion, electron microscopy serves as a standard method for morphological investigations of otoconia. Human utricular otoconia show a uniform outer morphology corresponding to a calcite-based nanocomposite. Morphological changes provide further evidence for degeneration of utricular otoconia in humans, which might be a preconditioning factor causing BPPV. In case of uncertain origin, particles can be clearly assigned to otoconial origin using EDX and XRD analyses.

Acoustically induced streaming flows near a model cod otolith and their potential implications for fish hearing.

The ears of fishes are remarkable sensors for the small acoustic disturbances associated with underwater sound. For example, each ear of the Atlantic cod (Gadus morhua) has three dense bony bodies (otoliths) surrounded by fluid and tissue, and detects sounds at frequencies from 30 to 500 Hz. Atlantic cod have also been shown to localize sounds. However, how their ears perform these functions is not fully understood. Steady streaming, or time-independent, flows near a 350% scale model Atlantic cod otolith immersed in a viscous fluid were studied to determine if these fluid flows contain acoustically relevant information that could be detected by the ear's sensory hair cells. The otolith was oscillated sinusoidally at various orientations at frequencies of 8-24 Hz, corresponding to an actual frequency range of 280-830 Hz. Phase-locked particle pathline visualizations of the resulting flows give velocity, vorticity, and rate of strain fields over a single plane of this mainly two-dimensional flow. Although the streaming flows contain acoustically relevant information, the displacements due to these flows are likely too small to explain Atlantic cod hearing abilities near threshold. The results, however, may suggest a possible mechanism for detection of ultrasound in some fish species.

Morphology and composition of Goldeye (Hiodontidae; Hiodon alosoides) otoliths.

We provide up-to-date morphological and compositional data on otoliths of the osteoglossomorph Goldeye (Hiodon alosoides). Using computed tomography (CT) X-ray, we documented the location of each of the three pairs of otoliths (lapilli, sagittae, and asterisci) in relation to the swim bladder, which extended forward in close proximity to the sagittae and asterisci. The lappili were the largest otoliths in terms of surface area and volume, but the sagittae were highly modified, appearing spiral in shape when viewed dorsally, with a surface area to volume ratio more than double that of the lapilli. Using scanning electron microscopy, the surface of each otolith was viewable in great detail, and small otoconia (~10.5 µm diameter) were observed on each, but were most numerous on the sagittae. On scanning electron micrographs, the sagittae appeared to be bi-lobed, with asymmetrical lobes each oriented in the same general direction. Using neutron and X-ray diffraction methods, we found three polymorphs of calcium carbonate crystals (aragonite, vaterite, and calcite), sometimes all within the same otolith. However, in general, lapilli and sagittae were composed predominately of aragonite whereas asterisci were composed chiefly of vaterite. With these results, we provide information on a unique species, whose inclusion in future studies would benefit our understanding of fish hearing, fish evolution, and fisheries ecology.

Analysis of Dix-Hallpike maneuver induced nystagmus based on virtual simulation.

BACKGROUND: How to interpret the various forms of nystagmus induced by the Dix-Hallpike maneuver has been the hotspot and difficulty of research. OBJECTIVES: Analysis of the types of nystagmus induced by Dix-Hallpike maneuver, and establish a diagnosis strategy based on dynamic nystagmus observation. MATERIALS AND METHODS: We observed the otolithic movements at different locations during the Dix-Hallpike maneuver through physical virtual simulation experiments and inferred the nystagmus performance, so as to establish the nystagmus interpretation rules for the repeated Dix-Hallpike maneuver. RESULTS: There are six types of nystagmus induced by the Dix-Hallpike maneuver. Nystagmus induced by the unilateral Dix-Hallpike maneuver does not accurately locate the otolith. The typical nystagmus that is consistent before and after the repetition of the Dix-Hallpike maneuver is the outward and upbeat nystagmus, considering the ipsilateral posterior semicircular canal BPPV. CONCLUSION: The atypical nystagmus often turns negative when the Dix-Hallpike maneuver is repeated. If the repeat test is positive and consistent with the results of the first diagnostic test, the otolith can be accurately located.

Evaluation of the structure of the otoconial layer using micro-computed tomography.

OBJECTIVE: Estrogen deficiency caused by bilateral ovariectomy (OVX) has been reported to lead to morphological changes in otoconia. Thus, we examined the morphological changes in the otoconial layer after OVX. We also investigated whether micro-computed tomography (µCT) is useful for the detection of morphological changes in the otoconial layer. METHODS: The otic capsules of C57BL/6 J mice were removed and evaluated using histological techniques and µCT at 2, 4, and 8 weeks after OVX or sham surgery. The volume of the utricle otoconial layer was measured and compared between the OVX and sham groups. The µCT scan and histological study results were also compared. RESULTS: The volume of the utricle otoconial layer was significantly increased 4 weeks after OVX compared to the sham group in both histological and µCT studies (p < 0.05). The volume of the otoconial layer measured using µCT was significantly correlated with the histological study results (p < 0.05). CONCLUSION: The volume of the utricle otoconial layer increased after OVX. These morphological changes could be detected by µCT.

Auditory chain reaction: Effects of sound pressure and particle motion on auditory structures in fishes.

Despite the diversity in fish auditory structures, it remains elusive how otolith morphology and swim bladder-inner ear (= otophysic) connections affect otolith motion and inner ear stimulation. A recent study visualized sound-induced otolith motion; but tank acoustics revealed a complex mixture of sound pressure and particle motion. To separate sound pressure and sound-induced particle motion, we constructed a transparent standing wave tube-like tank equipped with an inertial shaker at each end while using X-ray phase contrast imaging. Driving the shakers in phase resulted in maximised sound pressure at the tank centre, whereas particle motion was maximised when shakers were driven out of phase (180°). We studied the effects of two types of otophysic connections-i.e. the Weberian apparatus (Carassius auratus) and anterior swim bladder extensions contacting the inner ears (Etroplus canarensis)-on otolith motion when fish were subjected to a 200 Hz stimulus. Saccular otolith motion was more pronounced when the swim bladder walls oscillated under the maximised sound pressure condition. The otolith motion patterns mainly matched the orientation patterns of ciliary bundles on the sensory epithelia. Our setup enabled the characterization of the interplay between the auditory structures and provided first experimental evidence of how different types of otophysic connections affect otolith motion.

Investigation of inner ear anatomy in mouse using X-ray phase contrast tomography.

A thorough understanding of inner ear anatomy is important for investigators. However, investigation of the mouse inner ear is difficult due to the limitations of imaging techniques. X-ray phase contrast tomography increases contrast 100-1,000 times compared with conventional X-ray imaging. This study aimed to investigate inner ear anatomy in a fresh post-mortem mouse using X-ray phase contrast tomography and to provide a comprehensive atlas of microstructures with less tissue deformation. All experiments were performed in accordance with our institution's guidelines on the care and use of laboratory animals. A fresh mouse cadaver was scanned immediately after sacrifice using an inline phase contrast tomography system. Slice images were reconstructed using a filtered back-projection (FBP) algorithm. Standardized axial and coronal planes were adjusted with a multi-planar reconstruction method. Some three-dimensional (3D) objects were reconstructed by surface rendering. The characteristic features of microstructures, including otoconia masses of the saccular and utricular maculae, superior and inferior macula cribrosae, single canal, modiolus, and osseous spiral lamina, were described in detail. Spatial positions and relationships of the vestibular structures were exhibited in 3D views. This study investigated mouse inner ear anatomy and provided a standardized presentation of microstructures. In particular, otoconia masses were visualized in their natural status without contrast for the first time. The comprehensive anatomy atlas presented in this study provides an excellent reference for morphology studies of the inner ear.

Enigmatic ear stones: what we know about the functional role and evolution of fish otoliths.

Otoliths in bony fishes play an important role in the senses of balance and hearing. Otolith mass and shape are, among others, likely to be decisive factors influencing otolith motion and thus ear functioning. Yet our knowledge of how exactly these factors influence otolith motion is incomplete. In addition, experimental studies directly investigating the function of otoliths in the inner ear are scarce and yield partly conflicting results. Herein, we discuss questions and hypotheses on how otolith mass and shape, and the relationship between the sensory epithelium and overlying otolith, influence otolith motion. We discuss (i) the state-of-the-art knowledge regarding otolith function, (ii) gaps in knowledge that remain to be filled, and (iii) future approaches that may improve our understanding of the role of otoliths in ear functioning. We further link these functional questions to the evolution of solid teleost otoliths instead of numerous tiny otoconia as found in most other vertebrates. Until now, the selective forces and/or constraints driving the evolution of solid calcareous otoliths and their diversity in shape in teleosts are largely unknown. Based on a data set on the structure of otoliths and otoconia in more than 160 species covering the main vertebrate groups, we present a hypothetical framework for teleost otolith evolution. We suggest that the advent of solid otoliths may have initially been a selectively neutral 'by-product' of other key innovations during teleost evolution. The teleost-specific genome duplication event may have paved the way for diversification in otolith shape. Otolith shapes may have evolved along with the considerable diversity of, and improvements in, auditory abilities in teleost fishes. However, phenotypic plasticity may also play an important role in the creation of different otolith types, and different portions of the otolith may show different degrees of phenotypic plasticity. Future studies should thus adopt a phylogenetic perspective and apply comparative and methodologically integrative approaches, including fossil otoliths, when investigating otoconia/otolith evolution and their function in the inner ear.

Right-sided dominance of the bilateral vestibular system in the upper brainstem and thalamus.

MRI diffusion tensor imaging tractography was performed on the bilateral vestibular brainstem pathways, which run from the vestibular nuclei via the paramedian and posterolateral thalamic subnuclei to the parieto-insular vestibular cortex. Twenty-one right-handed healthy subjects participated. Quantitative analysis revealed a rope-ladder-like system of vestibular pathways in the brainstem with crossings at pontine and mesencephalic levels. Three structural types of right-left fiber distributions could be delineated: (1) evenly distributed pathways at the lower pontine level from the vestibular nuclei to the pontine crossing, (2) a moderate, pontomesencephalic right-sided lateralization between the pontine and mesencephalic crossings, and (3) a further increase of the right-sided lateralization above the mesencephalic crossing leading to the thalamic vestibular subnuclei. The increasing lateralization along the brainstem was the result of an asymmetric number of pontine and mesencephalic crossing fibers which was higher for left-to-right crossings. The dominance of the right vestibular meso-diencephalic circuitry in right-handers corresponds to the right-hemispheric dominance of the vestibular cortical network. The structural asymmetry apparent in the upper brainstem might be interpreted in relation to the different functions of the vestibular system depending on their anatomical level: a symmetrical sensorimotor reflex control of eye, head, and body mediated by the lower brainstem; a lateralized right-sided upper brainstem-thalamic function as part of the dominant right-sided cortical/subcortical vestibular system that enables a global percept of body motion and orientation in space.

Altered processing of otolithic information in isolated lateral medullary infarction.

Ocular and cervical vestibular-evoked myogenic potentials (VEMPs) evaluate the function of otolithic pathways in central as well as peripheral vestibular disorders. This study aimed to determine the associations and dissociations of otolithic dysfunction in lateral medullary infarction (LMI), the most well-known disorder of central vestibulopathy. At the Dizziness Clinic of a referral-based University Hospital, 45 patients with isolated LMI (28 men, mean age = 55.6 ± 12.5) had evaluation of the ocular tilt reaction (OTR), tilt of the subjective visual vertical (SVV), and ocular and cervical VEMPs from Janurary 2011 to August 2015 during the acute phase, 1-11 days from the symptom onset (median = 2 days). Almost all (42/45, 93 %) patients showed at least one component of the OTR or SVV tilt that was invariably ipsiversive. In contrast, oVEMPs were abnormal only in 12 (27 %) and cVEMPs in 13 (29 %) patients. Only three patients showed abnormal results in all the tests of the OTR, SVV tilt, and ocular and cervical VEMPs. Abnormal oVEMPs were more common in patients with the OTR than those without (38 vs 6 %, Pearson X 2 test, p = 0.021). In contrast, abnormality of cVEMPs showed no correlation with the presence of OTR (28 vs 31 %, Pearson X 2 test, p = 0.795). In patients with LMI, ipsiversive OTR is invariable, but abnormalities of oVEMPs and cVEMPs were much less common and mostly dissociated even in the patients with abnormal results. This discrepancy in otolithic dysfunction suggests different anatomical substrates and/or dissimilar reciprocal modulation for processing of each otolithic signal in central vestibular structures located in the dorsolateral medulla.

Ex vivo visualization of the mouse otoconial layer compared with micro-computed tomography.

HYPOTHESIS: Micro-computed tomography (micro-CT) is useful for assessing the 3-dimensional (3D) morphology and age-related changes of the otoconial layer. BACKGROUND: Wriggle mouse Sagami (WMS) is a mutant of the plasma membrane Ca2+ ATPase type2 gene (Atp2b2) with deficits in the saccular otoconia. We examined the effectiveness of micro-CT in evaluating the otoconial layer of WMS and C57BL/6J mice. METHODS: Otic capsules of C57BL/6J mice at different fixation time were examined using micro-CT to evaluate the effects of the fixation time on the otoconial layer. Otic capsules of Atp2b2(wri/wri), Atp2b2(wri/+), and Atp2b2(+/+) mice at P14, P21, and the age of 3 months were used to analyze age-related changes in the otoconial layer. A series of cross-section and 3D reconstructed images of the otoconial layer were obtained. The micro-CT findings were compared with the otic capsule findings cleared in methyl salicylate using stereomicroscopy. RESULTS: Micro-CT produced high-resolution images of the otoconial layer, thereby providing information regarding the spatial configuration and 3D curvature. There were no changes between the different fixation times. In the Atp2b2(wri/+) and Atp2b2(+/+) mice, the saccular and utricular otoconial layers were normal among all age groups. In the Atp2b2(wri/wri) mice, the saccular otoconial layer decreased on P14 and was absent on P21, whereas the utricular otoconial layer disappeared at 3 months of age. CONCLUSION: We obtained precise information regarding the mouse otoconial layer with minimum artifacts. This method is expected to improve our understanding of the physiologic function and age-related changes in otolith organs.

Computerized tomography of the otic capsule and otoliths in the oyster toadfish, Opsanus tau.

The neurocranium of the toadfish (Opsanus tau) exhibits a distinct translucent region in the otic capsule (OC) that may have functional significance for the auditory pathway. This study used ultrahigh resolution computerized tomography (100 µm voxels) to compare the relative density of three sites along the OC (dorsolateral, midlateral, and ventromedial) and two reference sites (dorsal: supraoccipital crest; ventral: parasphenoid bone) in the neurocranium. Higher attenuation occurs where structural density is greater; thus, we compared the X-ray attenuations measured, which provided a measure of relative density. The maximum attenuation value was recorded for each of the five sites (x and y) on consecutive sections throughout the OC and for each of the three calcareous otoliths associated with the sensory maculae (lagena, saccule, and utricle) in the OC. All three otoliths had higher attenuations than any sites in the neurocranium. Both dorsal and ventral reference sites (supraoccipital crest and parasphenoid bone, respectively) had attenuation levels consistent with calcified bone and had relatively small, irregular variations along the length of the OC in all individuals. The lowest relative attenuations (lowest densities) occurred consistently at the three sites along the OC. In addition, the lowest attenuations measured along the OC occurred at the ventromedial site around the saccular otolith for all seven fish. The decrease in bone density along the OC is consistent with the hypothesis that there is a low-density channel in the skull to facilitate transmission of acoustic stimuli to the auditory endorgans of the ear.

Crystallographic and chemical composition of otoconia in the salamander Pleurodeles waltl.

The aim of the present study was to define the morphology and the crystallographic and chemical composition of otoconia in different regions of the inner ear in Pleurodeles waltl (urodele amphibian). The inner ear of adults was microdissected and otoconia were analyzed by scanning electron microscopy (SEM), X-ray diffraction, energy dispersive X-ray (EDX) and transmission electron microscopy. Two types of crystals were detected by SEM. Otoconia had different shapes depending on their location in the membranous labyrinth. One type had a cylindrical body with a triplanar smooth facet at each end, the other ones had either a prismatic shape with flat sides and end faces or a fusiform shape with rounded body and pointed end. The forms corresponded to those previously identified by other authors. These two types of otoconia had different X-ray diffraction patterns. The cylindrical otoconia were calcitic and located in the utricle, the other ones were aragonitic and located in the saccule, lagena and endolymphatic sac. An analysis by EDX indicated that both types of otoconia contained about 95% calcium with trace quantities of sodium, magnesium, phosphorus, sulfur, chlorine and potassium. Trace amounts of strontium was only found in the aragonitic otoconia.

Non-invasive assessment of otolith formation during development of the Japanese red-bellied newt, Cynops pyrrhogaster.

Pre-mated adult female newts and embryos have been flown on the International Microgravity Laboratory-2 (IML-2) Space Shuttle flight in 1994 (Wiederhold et al., 1992b). With the specimens available from this flight, the calcification of otoliths, ulna, radius and backbone of the flown larvae and adult newts were analyzed. The experiments presented here studied the development of the otoliths on the ground. Otoliths of living newts, from embryo to adult, were observed in situ with the application of a new X-ray and bio-imaging analyzer system. For the establishment of this method, newts at different developmental stages were used. An imaging plate temporarily stores the X-ray energy pattern at the bio-imaging analyzer. A latent image on the imaging plate was transformed into a digital time series signal with an image reader. Acquired digital information was computed with the image processor. The processed information was recorded on film with an image recorder, in order to visualize it on an enlargement computed radiograph. To analyze development of the otoliths, photo-stimulated luminescence level was detected by an image analyzer, using transmitted X-ray photons. A single clump of otoconia could first be seen at stage 33. Stage-36 embryos first have distinguishable otoliths, with the utricle in front and saccule behind. Our results show that this X-ray method detects the otoliths equally as well as sectioning. In the newt, the mandibular/maxillary bone formed before the spine. It is suspected that for the newt embryo, living in water, feeding becomes necessary prior to support of the body.

The otoliths of the antarctic teleost Trematomus bernacchii: scanning electron microscopy and X-ray diffraction studies.

The authors studied the otoliths of the Nototheniid Trematomus bernacchii with scanning electron microscopy and X-ray diffraction analysis. Results obtained reveal that three otoliths are present: a large sagitta, a lapillus and a fragile asteriscus. Their sensorial faces appear finely decorated as shown by scanning electron microscopy (SEM). The sagitta and the lapillus are aragonitic while the asteriscus is vateritic, as demonstrated by X-ray diffraction.

Three-dimensional cone beam computed tomography imaging of the membranous labyrinth in patients with Meniere's disease.

CONCLUSION: Three-dimensional cone beam computed tomography (3DCT) images revealed characteristic malformations of the membranous labyrinth of the inner ear in Meniere's disease (MD). The morphology of the membranous region between the vestibular cecum of the cochlea and the saccule of ears with MD was compared to that of healthy ears. The present study supports the hypothesis proposed earlier that reuniting duct blockade is a result of the dislodgement of saccular otoconia. OBJECTIVE: To visualize the membranous labyrinth using 3DCT and to investigate the pathology of MD. METHODS: A preparatory study was conducted to determine the optimal 3DCT window settings for the detection of water, muscle, calcium carbonate (CaCO3), and bone. Based on this preparatory study, the ears of 13 healthy volunteers and 25 MD patients definitely diagnosed according to the criteria issued by the Committee on Hearing and Equilibrium of the American Academy of Otolaryngology, Head and Neck Surgery (AAO-HNS), were visualized. RESULTS: The differences in the membranous labyrinth between MD ears and healthy ears could be visualized using 3DCT. The images were classified into three types based on their morphological pattern. The ears of patients with MD were different from normal ears in terms of this classification.

Blockage pattern of longitudinal flow in Meniere's disease.

CONCLUSION: In the present study, classification of the patterns of 3D CT images of the ductus reuniens (reuniting duct) (RD), saccular duct (SD), and endolymphatic sinus (ES) gave more precise information for assessing the pathological condition of Meniere's disease (MD) than our previous study. OBJECTIVE: This study attempted to provide more detailed information on MD by classifying the patterns of 3D CT images of the RD, SD, and ES in patients with MD. METHODS: We examined the ears of 62 patients with definitely diagnosed unilateral MD based on the criteria of the Committee on Hearing and Equilibrium of the American Academy of Otolaryngology-Head and Neck Surgery (AAO-HNS) using 3D CT. The 3D CT images of bony grooves of RD, SD, and ES (BRD, BSD, and BES) were classified into patterns according to aspects of their patency. RESULTS: BRD could be classified into six types by assessing their patency defined using the criteria in this study. In the ears on the affected side of patients with MD, the BRD, BSD, and BES lost continuity in 3D CT images along their bony routes and were significantly different from normal healthy ears (p < 0.01). There were no significant differences among each stage of MD in the distributions of BRD and BES except for BSD.